Construction of gold-siRNA NPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana.

In recent years, gold nanoparticles (AuNPs) have been widely used as gene silencing agents and therapeutics for treatment of cancers due to their high transfection efficiency and lack of cytotoxicity, but their roles in gene silencing in plants have not yet been reported. Here, we report synthesis of AuNPs-branched polyethylenimine and its integration with the small interfering RNAs (siRNA) of NPR1 to form a AuNPs-siRNA NPR1 compound. Our results showed that AuNPs-siRNA NPR1 was capable of infiltrating into Arabidopsis cells. AuNPs-siRNA NPR1 silenced 80% of the NPR1 gene in Arabidopsis. Bacteriostatic and ion leakage experiments suggest that the NPR1 gene in Arabidopsis leaves was silenced by AuNPs-siRNA NPR1 . In Columbia-0 plants, compared with the control group treated with buffer solution, the AuNPs-siRNA NPR1 treatment significantly increased the number of colonies and cell death, and the leaves turned yellow, similar to the phenotype of the npr1 leaves. These results indicated this AuNPs-siRNA NPR1 silencing the NPR1 gene method is simple, effective and quick (3 days), and a powerful tool to study gene functions in plants.

Advantages with prophylactic PEG-rhG-CSF versus rhG-CSF in breast cancer patients receiving multiple cycles of myelosuppressive chemotherapy: an open-label, randomized, multicenter phase III study.

BACKGROUND: PEG-rhG-CSF reduces neutropenia and improves chemotherapy safety. In China's registration trial (CFDA: 2006L01305), we assessed its efficacy and safety against rhG-CSF, and prospectively explored its value over multiple cycles of chemotherapy. METHODS: In this open-label, randomized, multicenter phase 3 study, breast cancer patients (n = 569) were randomized to receive PEG-rhG-CSF 100 µg/kg, PEG-rhG-CSF 6 mg, or rhG-CSF 5 µg/kg/d after chemotherapy. The primary endpoints were the incidence and duration of grade 3/4 neutropenia during cycle 1. Secondary endpoints included the incidence and duration of grade 3/4 neutropenia during cycles 2-4, the incidence of febrile neutropenia, and the safety. RESULTS: A once-per-cycle PEG-rhG-CSF at either 100 µg/kg or 6 mg was not different from daily injections of rhG-CSF for either incidence or duration of grade 3/4 neutropenia. Interestingly, a substantial difference was noted during cycle 2, and the difference became bigger over cycles 3-4, reaching a statistical significance at cycle 4 in either incidence (P = 0.0309) or duration (P = 0.0289) favoring PEG-rhG-CSF. A significant trend toward a lower incidence of all-grade adverse events was noted at 129 (68.98%), 142 (75.53%), and 160 (82.47%) in the PEG-rhG-CSF 100 µg/kg and 6 mg and rhG-CSF groups, respectively (P = 0.0085). The corresponding incidence of grade 3/4 drug-related adverse events was 2/187 (1.07%), 1/188 (0.53%), and 8/194 (4.12%), respectively (P = 0.0477). Additionally, PFS in metastatic patients preferred PEG-rhG-CSF to rhG-CSF despite no significance observed by Kaplan-Meier analysis (n = 49, P = 0.153). CONCLUSIONS: PEG-rhG-CSF is a more convenient and safe formulation and a more effective prophylactic measure in breast cancer patients receiving multiple cycles of chemotherapy.

Association between the MTHFR C677T polymorphism and gastric cancer susceptibility: A meta-analysis of 5,757 cases and 8,501 controls.

Current data regarding the association between the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and the risk of developing gastric cancer are insufficient to draw definite conclusions. Therefore, the present meta-analysis was conducted to achieve a more precise estimation of the association. MEDLINE, EMBASE and Wanfang database searches resulted in the identification of 28 eligible studies describing 5,757 cases and 8,501 controls. The strength of the association between the MTHFR C677T polymorphism and gastric cancer risk were evaluated using crude odds ratios (ORs), with 95% confidence intervals (CIs). The pooled ORs were determined using homozygous (TT vs. CC), heterozygous (CT vs. CC), dominant (TT+CT vs. CC) and recessive (TT vs. CC+CT) models. When all studies were pooled into the meta-analysis, significant associations were identified between the MTHFR C677T polymorphism and the risk of gastric cancer (homozygous model: OR, 1.39; 95% CI, 1.20-1.62; heterozygous model: OR, 1.18; 95% CI, 1.05-1.32; dominant model: OR, 1.23; 95% CI, 1.10-1.38; recessive model: OR, 1.26; 95% CI, 1.12-1.42). Stratification of the data by ethnicity identified a statistically significantly elevated risk of gastric cancer in Asian MTHFR C677T polymorphism populations (homozygous model: OR, 1.64; 95% CI, 1.43-1.90; heterozygous model: OR, 1.30; 95% CI, 1.16-1.45; dominant model: OR, 1.39; 95% CI, 1.25-1.54; recessive model: OR, 1.41; 95% CI, 1.25-1.51), but not in Caucasian populations (homozygous model: OR, 1.15; 95% CI, 0.89-1.48; heterozygous model: OR, 1.03; 95% CI, 0.84-1.25; dominant model: OR, 1.05; 95% CI, 0.86-1.28; recessive model: OR, 1.09; 95% CI, 0.91-1.31). Following adjustment for heterogeneity, the current meta-analysis demonstrated that the MTHFR C677T polymorphism was not associated with the risk of gastric cancer in Caucasian individuals. Furthermore, no evidence of publication bias was observed. Thus, the current meta-analysis indicates that the MTHFR C677T allele may be a low-penetrant risk factor for the development of gastric cancer in Asian populations.

[Effect of Nogo-66 on retinal microglia in chronic ocular hypertension rats].

OBJECTIVE: To study the effect of protein Nogo-66 on the expression of CD11b and MHC-II in retinal microglia of chronic ocular hypertension SD rats and the effects of protein Nogo-66 on the immunogenicity of injured retinal tissues. METHODS: It was a control experimental study. Chronic ocular hypertension rat model was established by laser photocoagulation on the anterior chamber angle and superficial vein of the sclera. One ml of Nogo-66 (0.01%) in PBS was injected subcutaneously on the day of laser treatment and 0.005% Nogo-66 PBS solution was injected into the vitreous 7 days and 1 month latter. PBS without Nogo-66 was injected in the control group. The expression of cell surface antigen CD11b and MHC-II were detected by immunohistochemistry 1 month and 1 day after the establishment of hypertension model. The difference of average IOP among groups was analyzed by variance analysis. The difference of expression of CD11b and MHC-II between the experimental and control groups was analyzed by t-test. RESULTS: The intraocular pressure (IOP) of experimental groups rised from the seventh day after model-building and the highest IOP was (24.16 ± 2.70) mm Hg (1 mm Hg = 0.133 kPa) 1 month later while that in the control groups was (15.93 ± 3.28) mm Hg. The difference between them was statistically significant (F = 2.10, P < 0.05). Expression of CD11b was (1.78 ± 0.63)% and MHC-II was (3.92 ± 1.03)% in Nogo-66 with hypertension groups, these results was significantly lower than those in Nogo-66 with normal intraocular pressure groups in which the expression of CD11b was (8.15 ± 1.97)% (t = 2.35, P < 0.05) and MHC-II was (11.45 ± 1.97)% (t = 2.14, P < 0.05). CONCLUSIONS: Protein Nogo-66 activated the cell surface antigen CD11b in nerve fiber layer of retina and induced antigen presenting molecules (MHC-II). This indicates that Nogo has the center immunogenicity and this protein could activate antigen-presenting cells to present injury antigen.

A new staging system is more discriminant than conventional staging systems for unresectable hepatocellular carcinoma.

BACKGROUND: Prediction of the life expectancy of a patient with unresectable hepatocellular carcinoma (HCC) remains difficult. The aims of the study were to construct a new staging scheme for patients with unresectable HCC and to compare the discriminatory ability of the staging scheme with the Okuda and CLIP score and TNM staging system in a cohort of patients with unresectable HCC. METHODS: A retrospective analysis of unresectable HCC cases from 1999 to 2003 was performed. The Cox model was used for multivariate analyses. The final model was derived from 10 randomly chosen training samples and the prognostic validity of the new staging scheme was assessed on the corresponding testing samples. Moreover, 54 cases with unresectable HCC were enrolled and prospectively followed up. The new staging, named the China integrated score (CIS), Okuda, TNM and CLIP systems were determined for each case. Comparisons of the survival rate between each stage were performed to evaluate their discriminatory ability. RESULTS: A simple scoring system was constructed, assigning linear scores (0/1/2) to the three covariates (TNM, alpha-fetoprotein and Child-Pugh) of the final model. The CIS system was more discriminant than the Okuda or TNM staging system, as confirmed by the Kaplan-Meier comparison of survival curves and by the Cox's regression analysis, with a median survival rate of 9.0, 2.3, 2.1 and 0.6 months in patients with CIS 2, 3, 4 and 5, respectively. The CIS system was performed as well as the CLIP score. CONCLUSION: The new staging system, accounting for both liver function and tumor characteristics, can accurately identify patients with different prognoses, particularly in the advanced phases of HCC. It should be useful as the only tool that can be applied for patients with unresectable HCC.

[Construction of a therapeutic effect evaluation system for patients with primary liver cancer based on syndrome differentiation in traditional Chinese medicine].

OBJECTIVE: To construct a system of therapeutic effect evaluation for patients with primary liver cancer according to the theory of syndrome differentiation in traditional Chinese medicine (TCM), and to examine its reliability. METHODS: Analytic hierarchy process and 100 mm surveyor's rod method were applied to obtain bottom layer and top level syndromes, which were used to construct the method of therapeutic effect evaluation, and its reliability was verified in clinical practice by comparing with some evaluation criteria in Western medicine, such as cancer severity scale; Karnofsky performance scale; Child-Pugh classification, cancer staging classification, and quality of life scale, etc. RESULTS: A system of therapeutic effect evaluation was constructed, and it could reflect the progress of tumor, changes of hepatic function and constitution. The evaluation scores acquired from the system were highly associated with the quality of life of the patients. CONCLUSION: The system of therapeutic effect evaluation can reflect the severity of disease and the characteristics of TCM treatment.

[Clinical application of Lishengsu for treatment of leukopenia induced by chemotherapy].

The efficacy and safety of Lishengsu (a rhG-CSF preparation) were evaluated for treatment of chemotherapy-induced leukopenia. 327 cases of leukopenia with grade I-IV induced by chemotherapy were subcutaneously administered at 2.5 - 5.0 micro g/(kg x d) of Lishengsu, and hemogram and the side effects were observed. The results showed that Lishengsu had satisfactory effect to cure leukopenia after chemtherapy, with an effective rate of 99.4% (325/327), the side effects were quite slight. It is concluded that Lishengsu is efficient and safe for patients with leukopenia, and can be used as an adjuvant drug for treatment of leukopenia after chemotherapy.